2001
DOI: 10.1006/jmbi.2001.4914
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Escherichia coli maltose-binding protein as a molecular chaperone for recombinant intracellular cytoplasmic single-chain antibodies 1 1Edited by R. Huber

Abstract: Recombinant single-chain antibodies (scFvs) that are expressed in the cytoplasm of cells are of considerable biotechnological and therapeutic potential. However, the reducing environment of the cytoplasm inhibits the formation of the intradomain disul®de bonds that are essential for correct folding and functionality of these antibody fragments. Thus, scFvs expressed in the cytoplasm are mostly insoluble and inactive.Here, we describe a general approach for stabilizing scFvs for ef®cient functional expression i… Show more

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Cited by 166 publications
(128 citation statements)
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“…HP0149 (fused either to His 6 or MBP) was toxic; therefore, the interaction that HP0149 is engaged in could not be studied (Table II). These results are in agreement with several studies describing MBP tags as an efficient tag to enhance solubility of fusion proteins (33)(34)(35). Expression and solubility results of the His 6 tag proteins are overall comparable to those reported in other expression studies (35).…”
Section: Cloning Expression and Solubility Of The Proteinssupporting
confidence: 91%
“…HP0149 (fused either to His 6 or MBP) was toxic; therefore, the interaction that HP0149 is engaged in could not be studied (Table II). These results are in agreement with several studies describing MBP tags as an efficient tag to enhance solubility of fusion proteins (33)(34)(35). Expression and solubility results of the His 6 tag proteins are overall comparable to those reported in other expression studies (35).…”
Section: Cloning Expression and Solubility Of The Proteinssupporting
confidence: 91%
“…An unknown in these experiments is how fusion with MBP [used as a molecular chaperone to promote peptide solubility and stability (24)] might affect the presentation of the tilted peptide. MBP is a large protein with two distinct globular domains separated by a deep groove, and at its surface lie several clusters of hydrophobic residues (25).…”
Section: Discussionmentioning
confidence: 99%
“…40 Some of the examples given for scFvs that exhibited improved expression upon fusion to tags were already confirmed to be well-functioning intrabodies when expressed without a fusion tag. 40,48,49 Therefore, fusion to a Cκ-domain or to solubility enhancers might lead to enhanced expression or improved performance of an already confirmed functional intrabody, but this method may not be sufficient to convert all scFvs into intrabodies.…”
Section: ©2 0 1 1 L a N D E S B I O S C I E N C E D O N O T D I S Tmentioning
confidence: 99%